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Clathrin, adaptors and eps15 in endosomes containing activated epidermal growth factor receptors
Abstract
Activation of the epidermal growth factor receptor (EGFR) by EGF results in binding of clathrin adaptor protein complex AP-2 to the receptor cytoplasmic tail. The transient interaction with AP-2 is thought to be responsible for the selective recruitment of the EGFR into coated pits during endocytosis. In this study we found that EGF-induced EGFR/AP-2 association, measured by co-immunoprecipitation, persists after receptor internalization. Double-label immunofluorescence of EGF-treated A-431 and COS-1 cells revealed the presence of AP-2, clathrin and eps15, another component of the plasma membrane coated pits, in the large perinuclear endosomes loaded with EGFRs. By optical sectioning and image deconvolution, the immunoreactivities were seen to be distributed within vesicular and tubular elements of these endosomes. In addition, these compartments contained the transferrin receptors and a EEA.1 protein, markers of early endosomes. Furthermore, Golgi clathrin adaptor complex AP-1 was found in EGFR-containing endosomes and EGFR immunoprecipitates in A-431 cells. The direct interaction of the EGFR with µ1 as well as µ2 subunits of AP-1 and AP-2, correspondingly, was shown using the yeast two-hybrid assay. Brefeldin A, a drug that releases AP-1 from the trans-Golgi membranes, had no effect on AP-1 association with endosomes and its co-precipitation with EGFR. Taken together, the data suggest that endosomal EGFR-AP complexes make up a significant portion of the total amount of these complexes detectable by coimmunoprecipitation. It can be proposed that APs are capable of binding to the endosomal membrane via a mechanism that requires AP interaction with the intracellular tails of multimeric receptors like activated EGFR, which in turn allows recruitment of clathrin and eps15. The hypothesis that the competition between adaptor complexes for binding to the receptor tails in endosomes may regulate of the sorting of receptors is discussed.
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AP-1 μ1-adaptin can recognize the tyrosine-based motif Y974RAL and YXXΦ-independent interfaces in the cytoplasmic domains of EGFR.
“It has been reported that the μ1-adaptin subunit of AP-1 can recognize the tyrosine-based motif, Y974RAL, and YXXΦ-independent interfaces in the cytoplasmic domains of EGFR [34, 35]”
From Clathrin adapters AP-1 and GGA2 support expression of epidermal growth factor receptor for cell growth (2021) · cites this paper as [35]
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Metadata
- Authors
- Sorkina, T., Bild, A., Tebar, F., & Sorkin, A.
- Affiliations
- University of Colorado Health Sciences Center Department of Pharmacology , , 4200 E. Ninth Ave., Denver, CO 80262, USA
- Venue
- Journal of Cell Science
- Publisher
- The Company of Biologists
- Published
- 1999-02-01
- Volume
- 112
- Issue
- 3
- Pages
- 317-327
- ISSN
- 0021-9533
- 1477-9137
- Cited by
- 57
- References
- 0
- Language
- en
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